Simultaneous Hplc Determination Of 22 Components Of Essential Oils; Method Robustness With Experimental Design

958 words - 4 pages

The aim of the present study was the development and validation of a simple, precise and specific reversed phase HPLC method for the simultaneous determination of 22 components present in different essential oils namely cinnamon bark oil, caraway oil and cardamom fruit oil. The chromatographic separation of all the components was achieved on Wakosil–II C18 column with mixture of 30 mM ammonium acetate buffer (pH 4.7), methanol and acetonitrile in different ratio as mobile phase in a ternary linear gradient mode. The calibration graphs plotted with five different concentrations of each component were linear with a regression coefficient R2 > 0.999. The limit of detection and limit of quantitation were estimated for all the components. Effect on analytical responses by small and deliberate variation of critical factors was examined by robustness testing with Design of Experiment employing Central Composite Design and established that method was robust. The method was then validated for linearity, precision, accuracy, and specificity to demonstrate that it can be applicable to the determination of the ingredients in commercial sample of essential oils.

Key words: Reversed phase HPLC, cinnamon bark oil, caraway oil, cardamom fruit oil, Design of Experiment, essential oil, robustness testing.

Since time immemorial essential oils have been used for many purposes due to their diverse pharmacological properties. These are mainly complex mixtures of terpenoids and aromatic compounds. Cardamom oil is mainly used as a flavor, in flatulent indigestion and as an appetite stimulator. In addition it also possesses antibacterial, antiseptic, carminative and antispasmodic properties[1]. Caraway oil is used in pharmacy, perfumery, food and is also effective against spasmodic gastrointestinal complaints, flatulence, irritable stomach, indigestion, lack of appetite and dyspepsia in adults[2]. Cinnamon oil is used as a carminative, diuretic, antiflatulent, antipyretic, antiallergenic, anti-inflammatory and in cardiac disorders[3]. The sources of oils namely Ceylon cinnamon bark oil, cardamom fruit oil and caraway oil are Cinnamomum zeylancium Blume (synonym C. verum J Presl, Family: Lauraceae), Elettaria cardamomum Maton (Family: Zingiberaceae) and Carum carvi L (Family: Umbelliferae), respectively. Most of the reported chromatographic methods for quantitative determination of constituents of essential oils are mainly by GC or GC-MS. The main constituent of Ceylon cinnamon bark oil is trans cinnamaldehyde (72.0-82.15%), which is estimated by GC-MS analysis[4]. Several minor constituents generally present in all cinnamomum species viz. trans cinnamyl acetate (3.24-3.65%), eugenol (1.07-13.3%), tarns cinnamyl alcohol (0.5-0.6%), o-methoxy benzaldehyde (0.09-0.15%), benzyl benzoate (0.4-1.0%), α-terpineol (0.35-0.62%), β-caryophyllene (1.0-2.0%), linalool (0.7-1.06%), limonene (0.09-0.095%), myrcene (0.07-0.08%), β-pinene (0.02-0.03%), traces of...

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